CN105669787B - A method of extracting ring olivil 9- glucosides from Cortex Eucommiae - Google Patents
A method of extracting ring olivil 9- glucosides from Cortex Eucommiae Download PDFInfo
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- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
- C07H15/18—Acyclic radicals, substituted by carbocyclic rings
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- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
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- C07H1/08—Separation; Purification from natural products
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Abstract
A method of from 9 glucosides of Cortex Eucommiae extraction ring olivil, belonging to pharmaceutical technology field.The step of preparation method that ring olivil 4' glucosides is extracted in this Cortex Eucommiae, is followed successively by extraction, concentration, centrifugation, extraction, macroporous absorbent resin separation, silica gel column chromatography separation, the separation of ODS column chromatographies, the separation of MCI column chromatographies, the separation of 20 chromatographies of Sephadex LH, half prepares or preparative HPLC chromatographic isolation obtains 9 glucosides of ring olivil.Beneficial effects of the present invention：The present invention provides the extracting methods of 9 glucosides of Cortex Eucommiae middle ring olivil.This method has obtained 9 glucosides of ring olivil that purity is more than 98%.
Description
Technical field
The invention belongs to pharmaceutical technology fields, and in particular to Lignanoids compounds preparation method in Cortex Eucommiae.
Background technology
Cortex Eucommiae（Eucommia ulmoides Oliv.）For the dry bark of Eucommiaceae plant Cortex Eucommiae.Cortex Eucommiae is known as " living
Phytolite ", medicinal history is long,《Sheng Nong's herbal classic》Be classified as top grade, call its " cure mainly waist and knee, bowl spares, strengthening the essence gas,
Hard muscles and bones, except wet, dribbling urination of itching under the moon.Long term usage is made light of one's life by commiting suicide resistance to old." Cortex Eucommiae is the rare tonic Chinese herbal medicine of China's tradition, it is used for tonifying liver
Kidney, strengthening the bones and muscles.
See that the chemical composition of document report includes lignanoids, iridoids, Phenylpropanoid Glycosides class, flavonoids etc. in Cortex Eucommiae
About 200 chemical compositions altogether.Lignanoids compounds are most studies in chemical component of eucommia bark used, ingredient is most clear, structure is most clear
Clear a kind of compound, separated obtain is double tetrahydrofuran class compound mostly.So far, it is separated from Cortex Eucommiae
Lignanoids compounds to have 44, mostly glycosides compound, glycosyl be β-D-Glucose.Structure type is mostly bis-epoxy
Lignanoid（Bisepoxylignans, i.e. double tetrahydrofuran class）, such as Pinoresinol diglucoside, cloves fat diglucoside
Deng other also a small amount of monocycle oxygen lignanoid（monoepoxylignans）, neolignan（neolignans）, sesquilignan
（sesqiulignans）With ring lignanoid（cyclolignans）Deng.Have no the isolated ring olive resin from Cortex Eucommiae at present
The relevant report of plain 9- glucosides.
Invention content
The purpose of the present invention is according to defect described above, providing, a kind of recovery rate is high to extract ring olive resin from Cortex Eucommiae
The method of plain 9- glucosides.
Ring olivil 9- glucosides provided by the invention, has the following structure：
。
A kind of method of Cortex Eucommiae extraction ring olivil 9- glucosides, which is characterized in that pass sequentially through following steps preparation
It obtains：
（1）Cortex Eucommiae is extracted using the ethyl alcohol or methanol of 50%-95% through solvent extraction method, extracting solution is recovered under reduced pressure, obtains and slightly carries
Object；
（2）Take step（1）The crude extract of gained is centrifuged with being stood after distillation water dissolution, is taken supernatant；
（3）Take step（2）The supernatant petroleum ether extraction of gained discards petroleum ether extraction liquid, remaining aqueous solution decompression
It is concentrated to give medicinal extract；
（4）Take step（3）The medicinal extract of gained is detached through macroporous absorbent resin, with the ethanol elution of distilled water and 70%-95%,
Ethanol eluate is through being concentrated under reduced pressure to obtain ethanol elution object；
（5）Take step（4）The ethanol elution object of gained is detached through silica gel column chromatography, with chloroform/methanol mixed solvent ladder
Degree elution；
（6）Take step（5）Middle gained flow point is detached through ODS column chromatographies, with methanol/water or ethanol/water mixed solvent ladder
Degree elution；
（7）Take step（6）Middle gained flow point is detached through MCI column chromatographies, with methanol/water or ethanol/water mixed solvent ladder
Degree elution；
（8）Take step（7）Middle gained flow point is detached through Sephadex LH-20 chromatographies, mixed with methanol/water or ethanol/water
Bonding solvent elutes, and obtains ring olivil 9- glucosides crude products；
（9）Take step（8）Middle gained ring olivil 9- glucosides crude products are prepared through half or preparative HPLC chromatographic isolation,
With methanol/water, acetonitrile/water or methanol/acetonitrile/water elute for mobile phase, obtain ring olivil 9- glucosides.
Step in the preparation method of ring olivil 9- glucosides provided by the invention（3）Petroleum ether and supernatant volume ratio
It is 1:3~3:1, extraction times are 2~5 times.
Step in the preparation method of ring olivil 9- glucosides provided by the invention（4）When macroporous absorbent resin detaches,
The w/v of medicinal extract and distilled water is 1:20~1:30；The w/v of medicinal extract and ethyl alcohol is 1:10~1:40.
Step in the preparation method of ring olivil 9- glucosides provided by the invention（5）Eluting solvent is 1:0~20:1
Chloroform/methanol.
Step in the preparation method of ring olivil 9- glucosides provided by the invention（6）Eluting solvent is 1:9~4:0
Methanol/water or ethanol/water.
Step in the preparation method of ring olivil 9- glucosides provided by the invention（7）Eluting solvent is 1:10~4:0
Methanol/water or ethanol/water.
Step in the preparation method of ring olivil 9- glucosides provided by the invention（8）Eluting solvent is 1:10~2:1
Methanol/water or ethanol/water.
Step in the preparation method of ring olivil 9- glucosides provided by the invention（9）Mobile phase is methanol/water, second
Nitrile/water or methanol/acetonitrile/water are 1 from methanol/water mixed solvent ratio:10~1:0, acetonitrile/water mixed solvent ratio 1:20
~1:Ring olivil 9- glucosides is obtained in 0 flow point.
The beneficial effects of the invention are as follows：The present invention provides the extracting methods of Cortex Eucommiae middle ring olivil 9- glucosides.It should
Method has obtained the ring olivil 9- glucosides that purity is more than 98%, and recovery rate is high.
Specific implementation mode
The following examples will be further described the present invention, but not thereby limiting the invention.
Embodiment 1.
（1）Eucommia ulmoides 20kg, 95% ethyl alcohol being separately added into, heating and refluxing extraction 3 times 2 hours every time, are recovered under reduced pressure
Extracting solution obtains alcohol extracts.
（2）Alcohol extracts add 50L distilled water, and 12 hours are stood after dissolving, and it is viscous to obtain brown after rotating speed 5000 rpm centrifugations
Thick precipitation and clarified supernatant, tan precipitate discard.
（3）To step（2）Isometric petroleum ether is added in middle clarified supernatant, after fully stirring evenly, stands 12 hours, takes out
Supernatant, then isometric petroleum ether is added into remaining clear solution, repetition aforesaid operations are secondary, and petroleum ether extraction liquid is abandoned
It goes.Remaining clear solution is concentrated under reduced pressure, and obtains medicinal extract 2.2kg.
（4）Take step（3）Middle medicinal extract 2.2kg is detached through macroporous absorbent resin HPD 100, distills water elution, distillation with 50L
Water elution discards.Then with 60 L, 95% ethanol elutions, eluent obtains ethanol elution object after reduced pressure.
（5）Take step（4）Middle ethanol elution object, detaches through silica gel column chromatography, uses chloroform：Methanol is respectively 1:0,20:
1,10:1,7:1,5:1,2:1,1:1 carries out gradient elution.
（6）Take step（5）In with chloroform：Methanol is 7:1 elution gained flow point, detaches through ODS column chromatographies, uses methanol：
Water is respectively 1:9,1:4,2:3,3:2,4:1,1:0 carries out gradient elution.
（7）Take step（6）Middle methanol：Water is 1:4 elution gained flow points, detach through MCI column chromatographies, use methanol：Moisture
It Wei 1:9,1:4,2:3,3:2,4:1,1:0 carries out gradient elution.
（8）Take step（7）Middle methanol：Water is 2:3 elution gained flow points, detach through Sephadex LH-20 column chromatographies,
Use methanol：Water is 3:7 are eluted, and ring olivil 9- glucosides crude products are obtained.
（9）Ring olivil 9- glucosides crude product is through preparing HPLC chromatogram separation, with methanol：Water is 28:72 isocratic elutions,
Flow velocity is 2ml/min, and ring olivil 4'- glucosides 2.15g, purity 98.3% are obtained within 18 minutes in retention time.
The Structural Identification data of ring olivil 9- glucosides are as follows：
White powder, Molisch reactions are positive, and sour water solution detects D-Glucose.IR (KBr) cm-1:3391,
2936,1605,1514,1450,1371,1256,1155,1076,1031,861,775,630,580,533. UV λmax
(CH3OH) nm:207,284.HR ESI MSm/z: 561.1955 ([M+Na]+, C26H34NaO12, Calcd
561.1948), 573.1742 ([M+Cl]-, C26H34ClO12, Calcd 573.1739)。
1H NMR ( 600 MHz, MeOD) δ: 6.80 ( 1H, d, J =1.9 Hz, H-2), 6.76 (1H,
d, J =7.9 Hz, H-5), 6.66 (1H, dd,J =1.9,7.9 Hz, H-6), 6.63 (1H, 3, H-2'
), 6.18 (H, s, H-5'), 4.12-4.15 (2H, m, 7,9-H), 4.07 (1H, d,J = 7.9 Hz, H-
1''), 3.85 (1H, d,J =11.3 Hz, H-9'), 3.81 (6H, s, 3,3'-OCH3 ), 3.54 (1H,
d, J =11.3 Hz, H-9'), 3.45 (1H, dd,J =4.3,10.6 Hz, H-9), 3.26 (1H, d,J
=16.6 Hz, H-7'), 2.61 (1H, d,J = 16.6 Hz, H-7' )。
13C NMR ( 150 MHz, MeOD) δ:149.1 (C-3), 147.5 (C-3'), 146.1 (C-4
), 145.2 (C-4'), 138.2 (C-1), 133.5 (C-1'), 126.5 (C-6'), 123.4 (C- 6),
117.3 (C-5'), 116.2 (C-5), 114.4 (C-2), 113.0 (C-2'), 105.3 (C-1 "), 78.1
(C-3 "), 78.0 (C-5''), 75.1 (C-2''), 74.5 (C-8'), 71.5 (C-4''), 69.2 (C-9
), 68.8 (C-9'), 62.6 (C-6''), 56.5 (- OCH3 ), 56.4 (- OCH3 ), 46.7 (C-8), 44.9
(C-7), 40.0 (C-7').
Embodiment 2.
（1）Eucommia ulmoides 100kg, 85% ethyl alcohol being separately added into, heating and refluxing extraction 3 times 2 hours every time, are recovered under reduced pressure
Extracting solution obtains alcohol extracts.
（2）Alcohol extracts add 250L distilled water, and 12 hours are stood after dissolving, and it is viscous to obtain brown after rotating speed 5000 rpm centrifugations
Thick precipitation and clarified supernatant, tan precipitate discard.
（3）To step（2）2 times of volume petroleum ethers are added in middle clarified supernatant, after fully stirring evenly, stands 12 hours, takes
Go out supernatant, then isometric petroleum ether is added into remaining clear solution, repetition aforesaid operations are secondary, and petroleum ether extraction liquid is abandoned
It goes.Remaining clear solution is concentrated under reduced pressure, and obtains medicinal extract 10.5kg.
（4）Take step（3）Middle medicinal extract 10.5kg is detached through macroporous absorbent resin HPD 100, is distilled water elution with 300L, is steamed
Distilled water eluent discards.Then with 400 L, 95% ethanol elutions, eluent obtains ethanol elution object after reduced pressure.
（5）Take step（4）Middle ethanol elution object, detaches through silica gel column chromatography, uses chloroform：Methanol is respectively 20:1,7:
1,3:1,1:1 carries out gradient elution.
（6）Take step（5）In with chloroform：Methanol is 7:1 elution gained flow point, detaches through ODS column chromatographies, uses methanol：
Water is respectively 1:9,1:4,2:3,4:1 carries out gradient elution.
（7）Take step（6）Middle methanol：Water is 1:4 elution gained flow points, detach through MCI column chromatographies, use methanol：Moisture
It Wei 1:4,2:3,3:2,1:0 carries out gradient elution.
（8）Take step（7）Middle methanol：Water is 2:3 elution gained flow points, detach through Sephadex LH-20 column chromatographies,
Use ethyl alcohol：Water is 2:3 are eluted, and ring olivil 9- glucosides crude products are obtained.
（9）Ring olivil 9- glucosides crude product is through preparing HPLC chromatogram separation, with acetonitrile：Water is 20:80 isocratic elutions,
Flow velocity is 2ml/min, and ring olivil 9- glucosides 11.07g, purity 99.4% are obtained within 25 minutes in retention time.
The Structural Identification method of ring olivil 9- glucosides is the same as embodiment 1.
Claims (8)
1. a kind of method of Cortex Eucommiae extraction ring olivil 9- glucosides, which is characterized in that pass sequentially through following steps and be prepared into
It arrives：
（1）Cortex Eucommiae is extracted using the ethyl alcohol or methanol of 50%-95% through solvent extraction method, extracting solution is recovered under reduced pressure, obtains crude extract；
（2）Take step（1）The crude extract of gained is centrifuged with being stood after distillation water dissolution, is taken supernatant；
（3）Take step（2）The supernatant petroleum ether extraction of gained, discards petroleum ether extraction liquid, and remaining aqueous solution is concentrated under reduced pressure
Obtain medicinal extract；
（4）Take step（3）The medicinal extract of gained is detached through macroporous absorbent resin, with the ethanol elution of distilled water and 70%-95%, ethyl alcohol
Eluent is through being concentrated under reduced pressure to obtain ethanol elution object；
（5）Take step（4）The ethanol elution object of gained is detached through silica gel column chromatography, is washed with chloroform/methanol mixed solvent gradient
It is de-；
（6）Take step（5）Middle gained flow point is detached through ODS column chromatographies, is washed with methanol/water or ethanol/water mixed solvent gradient
It is de-；
（7）Take step（6）Middle gained flow point is detached through MCI column chromatographies, is washed with methanol/water or ethanol/water mixed solvent gradient
It is de-；
（8）Take step（7）Middle gained flow point is detached through Sephadex LH-20 chromatographies, molten with methanol/water or ethanol/water mixing
Agent elutes, and obtains ring olivil 9- glucosides crude products；
（9）Take step（8）Middle gained ring olivil 9- glucosides crude products are prepared through half or preparative HPLC chromatographic isolation, with first
Alcohol/water, acetonitrile/water or methanol/acetonitrile/water elute for mobile phase, obtain ring olivil 9- glucosides.
2. a kind of method of Cortex Eucommiae extraction ring olivil 9- glucosides according to claim 1, it is characterised in that：Step
（3）Petrochina ether is 1 with supernatant volume ratio:3~3:1, extraction times are 2~5 times.
3. a kind of method of Cortex Eucommiae extraction ring olivil 9- glucosides according to claim 1, it is characterised in that：Step
（4）When middle macroporous absorbent resin separation, the w/v of medicinal extract and distilled water is 1:20~1:30；The weight of medicinal extract and ethyl alcohol
Volume ratio is 1:10~1:40.
4. a kind of method of Cortex Eucommiae extraction ring olivil 9- glucosides according to claim 1, it is characterised in that：Step
（5）Middle eluting solvent is 1:0~20:1 chloroform/methanol.
5. a kind of method of Cortex Eucommiae extraction ring olivil 9- glucosides according to claim 1, it is characterised in that：Step
（6）Middle eluting solvent is 1:9~4:0 methanol/water or ethanol/water.
6. a kind of method of Cortex Eucommiae extraction ring olivil 9- glucosides according to claim 1, it is characterised in that：Step
（7）Middle eluting solvent is 1:10~4:0 methanol/water or ethanol/water.
7. a kind of method of Cortex Eucommiae extraction ring olivil 9- glucosides according to claim 1, it is characterised in that：Step
（8）Middle eluting solvent is 1:10~2:1 methanol/water or ethanol/water.
8. a kind of method of Cortex Eucommiae extraction ring olivil 9- glucosides according to claim 1, it is characterised in that：Step
（9）Middle mobile phase is methanol/water, acetonitrile/water or methanol/acetonitrile/water, and methanol/water mixed solvent ratio is 1:10~1:0, second
Nitrile/water mixed solvent ratio 1:20~1:0.
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| CN102526165A (en) * | 2010-12-07 | 2012-07-04 | 中国医学科学院药物研究所 | Rhodiola effective fractions, preparation method, drug composition and uses thereof |
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| CN102526165A (en) * | 2010-12-07 | 2012-07-04 | 中国医学科学院药物研究所 | Rhodiola effective fractions, preparation method, drug composition and uses thereof |
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| Title |
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| Chemical constituents fromStauntonia brachyanthera Hand–Mazz;Chao Chen et al.;《Biochemical Systematics and Ecology》;20130124;第48卷;182-185 * |
| New Chemical Constituents of Roots of Urtica triangularis HAND-MASS;Xing-Guo YAN et al.;《Chem. Pharm. Bull.》;20081031;第56卷(第10期);1463-1465 * |
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